ObjectiveTo determine whether transplanting olfactory ensheathing cells(OECs)is effective in controlling or re.versing the deterioration caused by amyotrophic lateral sclerosis(AtS). MethodsUetwcen February 2003 and April 2006,327 pa-fients(241 males and 86 females)with probable or definite ALS(diagnosed according to the El Escorial criteria)received dle oECstransplantation.Their ages ranged from 20 to 84 years(51.6±11.1 years).The duaration of sympltoms before surgical trealment wit84.8months to 13 years(2.9±2.0 years).OECs were cultured and.injected into palllological regions of the spinal cord and/or bilateralcoroila radiata of the brain;the patients were divided into three groups,group A(cord only,n=29),group B(cord and brain,,n=6),and group C(brain only,n=292)based on the transplant sites.ResultsThe patient's neurological function was assessedboth before and at4 weeks after transplantation by using the Amyolrophic Lateral Sclerosis Functional Rating Scale(ALSFRS)of the ALSCNTF Trealment Study(ACTS).The$cores were increased from 17.2±8.6 pre-operation to 20.1±9.7 post-operation in group A(P<0.05),from 24.2 4-6.8 to 25.7±6.6(P>0.05)in group B,and from 20.3±8.6 to 22.0±9.4(P<0.001)in group C.There were no significant difference inincreased ALSFRS scores amongthe three groups(P>O.05).The total improvement rate of neurological function was 77.1%(252/327).The result of electramyographic examination showed that spontaneous potential diminishedand/or disappeared,the amplitude of the motor unit action potential decreased remarkably andthe numbers of motor unitaction potentialgreatly increased in 261 cases(79.8%).Sixteen patients(4.9%)experienced the various complications including headache.short-term fever,seizure attack,central nerve system infection,pneumonia,respiratory failure,urinary tract infection,heartfailure,and pos-sible pulmonary embolism;of them,there were 4 deaths(1.2%). ConclusionThese preliminary results suggest that the OECs trasplantafion is effective in controlling or reversing the physiological deterioration caused by ALS.

Objective To evaluate the safety and efficacy of one-level posterior lumbar interbody fusion(PLIF) combined with Prospace and facet fusion using local autograft. Methods Clinical and radiographic data of 76 patients treated by this technique was reviewed from May 2002 to December 2004. Of them, there were 52 males and 24 females, with an average age of 53.2 years (23-81 years), including 60 cases of degenerative disc disease, 9 cases of failed back surgery syndrome and 3 cases of spondylolysis. The disese courses were 1.2-8.7 years (mean 3.6 years). The levels of PLIF were: L2,3 in 2 cases, L3,4 in 7, L4,5 in 54, L5/S1 in 10, L4/S1 in 1 and L5,6 in 2. After decompression, Prospace was inserted into interbody space bilaterally,and located in disc space 4 mm beyond the rear edge of the vertebral body. Local laminectomy autograft was packed both laterally into and between 2 implants. Then the remanent local autograft was placed over facet bed. Pedicle screws were used after insertion of Prospace. Clinical results were evaluated by the JOA score. Disc height ratio and lumbar lordosis angles were measured on lateral radiographs. Fusion status was determined by evidence of bridge trabeculae across facet joint and interbody space on CT scan without mobility in lateral dynamic X-rays, and no radiolucent gap between Prospace and endplate. Paired t-test was used for statistical analysis. Results Mean blood loss and operative time was 384 ml and 178 minutes, respectively. The average JOA score at final follow-up (26.1 + 2.7) was significantly improved when compared with that of pre-operation (14.5 ± 4.0, P ＜ 0.05), with a mean recovery rate of JOA score 81.1% (37.5%-100.0%). The fusion rate was 97.4% (74/76). Mean disc height ratio and the involved segmental lordosis angle were increased from preoperative 0.27 ±0.07 and 5.8 + 2.2° to 0.33 + 0.06 and 11.3 + 2.0° respeetively at the final follow-up, and the differences were significant ( P ＜ 0.05). There were no device-related complications. Conclusion This surgical technique combined with Prospace interbody device is a safe and effective surgical option for patients with one-level lumbar disorders when PLIF is warranted.

Objective To study the clinical effects of the artificial vertebral body of the biomimetic nano-hydroxyapatite/polyamide 66 (n-HA/PA66) composite for the structural reconstruction and the height restoring of the vertebral body in the thoracolumbar fractures by the anterior surgical procedures. Methods From December 2003 to January 2006, 42 patients with thoracolumbar fractures received the anterior surgical procedures to decompress and reconstruct the spinal vertebral structure with the artificial vertebral body of the n-HA/PA66 composite. Among the patients, there were 28 males and 14 females, aged 17-67 years, averaged 43.6 years. The thoracolumbar fractures developed at T12 in 5 patients, at L1 in 17, at L2 in 14, and at L3 in 6. The height of the anterior border of the vertebral body amounted to 29%-47% of the vertebral body height, averaged 40.6%. The Cobb angle on the sagittal plane was 21-38° averaged 27.6°. According to the Frankel grading scale, the injuries to the nerves were as the following: Grade A in 7 patients, Grade B in 19, Grade C in 8, Grade D in 6, and Grade E in 2. Results All the 42 patients were followed up for 6-25 months. Among the patients, 36 were reconstructed almost based on the normal anatomic structure, and 6 were well reconstructed. The mean height of the anterior border of the vertebral body was 40.6% of the vertebral body height before operation but 91.7% after operation. And the reconstructed height of the vertebra was maintained. The mean Cobb angle on the sagittal plane was 27.6°before operation but 13.4° after operation. All the patients had a recovery of the neurological function that had a 1-grade or 2-grade improvement except 7 patients who were still in Grade A and 2 patients who were in Grade D. The implant was fused 3-5 months after operation. No infection, nail break, bar/plate break or loosening of the internal fixation occurred. Conclusion The artificial vertebral body of the biomimetic n-HA/PA66composite can effectively restore the height and the structure of the vertebra, can be fused with the vertebral body to reconstruct the spinal structural stability effectively, and can be extensively used in the clinical practice.

Objective To evaluated the role of wt-P53 protein in telomerase regulation in keloid fibroblasts(KFBs). Methods The fibroblasts were derived from human keloid tissue which was proved by pathological diagnosis. KFBs were divided into 2 groups, the transfection group and the untransfection group. wt-p53 gene was transfected into the fibroblasts by adenovirus vectors in the transfection group. The KFBs untransfected with wt-p53 gene served as control (untransfection group). After 48 hours of transfection, the expression of wt-P53 protein was analyzed by both Western blotting and immunofluorescence method, respectively. The telomerase activity was evaluated by TRAP-ELISA after 1-7 days of transfection.Results All the KFBs from 2 groups expressed wt-P53 protein. But the expression level of wt-P53 protein in the transfection group was significantly higher than that in the untransfection group. At the same time of high expression of wt-P53 protein, the telomerase activity of KFBs in transfection group was significantly lower than that in the untransfection group( P＜0.05). Conclusion High level expression of wt-P53 protein can transiently inhibit the telomerase activity of KFBs.

Objective To review and summarize the latest development of the therapy for the Duchenne muscular dystrophy (DMD). Methods The recently-published articles related to the therapies for DMD were extensively reviewed and briefly summarized. Results The therapeutic approaches for DMD included the gene therapy, the cell therapy, and the pharmacological therapy.The gene therapy and the cell therapy were focused on the treatment for the cause of DMD by the delivery of the missing gene, the modification of the mutated gene, and the transfer of the normal cells including the stem cells, while the pharmacological therapy dealt with the downstream events caused by the dystrophin gene defect, slowed down the pathologic progress of DMD, and improved the DMD patient's life quality and life span, by medication and other factor treatments. Conclusion There is still no cure for DMD because of various difficulties in replacing or repairing the defected gene and of the multifaceted nature of the severe symptoms. Therefore, it is imperative for us to find out a more effective treatment that can solve these problems.

Objective To evaluate proton MR spectroscopy (1H-MRS) for detection of the motor cortex and adjacent brain in amyotrophic lateral sclerosis (ALS) patients with apparent upper motor neuron involvement after olfactory ensheathing cells(OECs) transplantation. Methods From December 2004 to February 2005, 7 patients with clinically definite ALS who could safely undergo MRS were admitted into the perspective study. The neurological status, ALS functional rating scale (ALSFRS), EMG, and 1H-MRS taken before and 2 weeks after operations were carefully analyzed. The NAA/Cr and Cho/Cr ratios were measured in the cerebral peduncle, genu and posterior limb of the internal capsule, corona radiata and precentral gyrus. Results The ALSFRS in 2 cases improved obviously whose ALSFRS increased from 30 to 33 and from 29 to 34 respectively. And 5 cases remained stable 2 weeks after OECs transplantation. Statistical analyses for all seven cases showed both the NAA/Cr and Cho/Cr ratios decreased, but in the two cases with ALSFRS improvement the NAA/Cr increased in the certain anatomic position which confirmed the neurological and EMG findings. Conclusion The proton MR spectroscopy is a suitable noninvasive measure for ALS evaluation. The preliminary study suggests that two of the seven ALS cases improved apparently short-term after OECs transplantation. More patients are required for the clinical study and longer follow-up duration is needed for future research.

Objective To investigate the therapeutic effects of the vacularized iliac graft for ischemic necrosis of the femoral head in Niger young patients with sickle cell disease. Methods From November 1998 to Apirl 2001, 12 patients(5 males and 7 females, aging 11-22 years) with sickle cell disease suffered from ischemic necrosis of the femoral head in 14 hips. The lesion was on one hip in 10 patients and on bilateral hip in 2 patients. Necrosis was classified as Ficat Stage Ⅲ-Ⅳ in all patients. Twelve hips in 12 patients were treated with vascularized iliac graft in Maradi Province Hospital of Niger. The effects were evaluated on the basis of the Harris hip score and radiological examination. Results The patients were followed up 24-30 months after operation(27.4 months on average). The clinical results were satisfactory though no radiological evidence of improvement was noted. The mean Harris hip score increased from 75 points to 90 points. Pain relief and hip function improvement were achieved in all patients. No patients needed revision during short-term follow-up of 24-30 months. Conclusion The method is recommendation for treatment of ischemic necrosis of the femoral head in the Ficat Stages Ⅲ- Ⅳ in young patients with sickle cell disease for pain relief and restoration of hip joint function. This procedure may prolong the timing and acting of total hip arthroplasty.

Objective To determine the safety of the fetal olfactory ensheathing cell(OEC) transplantation in patients with chronic spinal cord injury (SCI) by examination of the magnetic resonance imaging (MRI). Methods A prospective clinical study involving 16 patients with chronic SCI was designed to investigate the feasibility and biological safety of the fetal OEC transplantation in treatment of SCI. The olfactory bulbs from the 3-4-month-old aborted human fetuses following the strict ethical guidelines were harvested and trypsinized down to single fetal OEC. These cells were then cultured for 12-17 days and were prepared for a clinical use. From November 2001 to December 2002, 16 patients with chronic SCI were randomly enrolled. The patients suffered from SCI for 1.5-8 years (average 4.3 years) after the injury. The suspension (50 μl) containing about 1×106 fetal OECs was transplanted by an injection into the patients' spinal cords above and below the injury site. All the patients were assessed before the transplantation and were followed up with MRI for 29-42 months (average 38 mon) after the transplantation. Results No cell-related adverse effects were observed in any patient during the follow-up period. The follow-up with MRI did not reveal any development of optic glial tumor, tumor-like mass, new hemorrhage, edema, expanding cyst, new cyst formation, infection or disruption of the neural structure in the transplant site of all the patients. Conclusion This is the first clinical study demonstrating the long-term safety of the OEC therapy for SCI. The results indicate that our protocol is feasible and safe in treatment of patients with chronic SCI within 38 months after the injury. Although the size of the samples for our study was not big enough, the positive results of the study have encouraged us to make a further research in this field.

Objective To explore the influence factors for the functional improvement after the fetal olfactory ensheathing cell (OEC) transplantation for chronic spinal cord injury(SCI). Methods The olfactory bulbs were harvested and trypsinized down to single fetal OEC. They were cultured for 12-17 days prepared for use. From November 2001 to December 2003, a total of 300 patients volunteered for the fetal OEC transplantation, among whom 222 suffered from complete chronic SCI and 78 suffered from incomplete chronic SCI. The procedures were performed on the patients with a disease course ranging from 6 months to 31 years (average 3.1 years) after their injuries. The fetal OEC was transplanted by the form of injections into the spinal cord at the upper and lower ends of the injury site. All the patients were assessed by the ASIA standard before the transplantation and 2-8 weeks after the transplantation. The influence factors including age, sex, duration after the injury, and injury degrees and levels were compared with those in the functional improvement after fetal OEC transplantation. Results The partially-improved neurological functions assessed by the ASIA standard were indicated by the motor scores increasing from 39.1±20.6 to 45.9±20.3 (P<0.001), the light touch scores from 51.7±24.9 to 63.4±23.0 (P<0.001), and the pin prick scores from 53.0±24.2 to 65.3±22.7(P<0.001). There was no significant difference in the functional improvement of the motor, light touch, and pin brick when compared with the age, sex, duration after the injury, and the injury degrees and levels. The motor scores and light touch scores at the cervical level were higher than the scores at the thoracic level. Conclusion The fetal OEC transplantation can partially improve the neurological functions quickly in treatment of the chronic spinal cord injury. All the influence factors except the motor scores and light touch scores, which were higher at the cervical level than at thoracic level, have no impact on the functional improvement after the fetal OEC transplantation.

Objecttive To investigate the possibility of repairing articular cartilage defects with the mesenchymal stem cells (MSCs)- seeded type Ⅰ collagen-glycosaminoglycan(CG) matrices after being cultured with the chondrogenic differentiation medium.Methods The adherent population of MSCs from bone marrow of 10 adult dogs were expanded in number to the 3rd passage. MSCs were seeded into the dehydrothennal treatment (DHT) cross-linked CG matrices; 2 × 106 cells per 9-mm diameter samples were taken.Chondrogenic differentiation was achieved by the induction media for 3 weeks. Cell contractility was evaluated by the measuement of the cell-mediated contraction of the CG matrices with time in culture. The in vitro formation of the cartilage was assessed by an assay employing immunohistochemical identification of type Ⅱ collagen and by immunohistochemistry to demonstrate smooth muscle actin (SMA).The cells seededing CGs were implanted into cartilage defects of canine knee joints. Twelve weeks after surgery, the dogs were sacrificed and results were observed. Results There was significant contraction of the MSCs-seeded DHT cross-linked CG scaffolds cultured in the cartilage induction medium. After 21 days, the MSC-seeded DHT cross-linked matrices were contracted to 64.4% ± 0.3%; histologically, the pores were fotmd to be compressed and the contraction coupled with the newly synthesized matrix, transforming the MSCsseeded CG matrix into a solid tissue in most areas. The type Ⅱ collagen staining was positive. The SMA staining was positive when these MSCs were seeded and the contracted CGs were implanted into the cartilage defects of the canine knee joints to repair the cartilage defects. The function of the knee joints recovered and the solid cartilaginous tissue filled the cartilage defects. Conclusion The results demonstrates that MSCs grown in the CG matrices can produce a solid cartilaginous tissue containing type Ⅱ collagen after being cultured with the chondrogenic differentiation medium and implanted into cartilage defects. We hypothesize that the following steps can be performed in the chondrogenic process: ①MSCs express SMA, resulting in matrix contraction, thus achieving a required cell density (allowing the cells to operate in a necessary society); ②Cells interact to form a type Ⅱ collagen-containing extracellular matrix (and cartilaginous tissue); ④Other factors, such as an applied mechanical stress, may be required to form a mature cartilage with the normal architecture.